Niacin conjugated fatty acid mixtures and their uses

ABSTRACT

The invention relates to niacin conjugated fatty acid mixtures; compositions comprising an effective amount of a niacin conjugated fatty acid mixture; and methods for treating or preventing an metabolic disorder comprising the administration of as effective amount of a niacin conjugated fatty acid mixture.

PRIORITY

This application is related to U.S. Provisional Application No.61/238,903, filed Sep. 1, 2009, U.S. Provisional Application No.61/308,524, filed Feb. 26, 2010, and U.S. Provisional Application No.61/310,952, filed Mar. 10, 2010, and claims the benefit of U.S. Ser. No.12/872,555 filed Aug. 30, 2010. The entire disclosures of thoseapplications are relied on and incorporated into this application byreference.

FIELD OF THE INVENTION

The invention relates to fatty acid niacin conjugates; compositionscomprising an effective amount of a fatty acid niacin conjugate; andmethods for treating or preventing a metabolic disease comprising theadministration of an effective amount of a fatty acid niacin conjugate.The invention also relates to niacin conjugated fatty acid mixtures;compositions comprising an effective amount of a niacin conjugated fattyacid mixture; methods of producing niacin conjugated fatty acidmixtures, and methods for treating or preventing a metabolic disordercomprising the administration of an effective amount of a niacinconjugated fatty acid mixture. All patents, patent applications, andpublications cited herein are hereby incorporated by reference in theirentireties.

BACKGROUND OF THE INVENTION

Oily cold water fish, such as salmon, trout, herring, and tuna are thesource of dietary marine omega-3 fatty acids, eicosapentaenoic acid(EPA) and docosahexaenoic acid (DHA) being the key marine derivedomega-3 fatty acids. Both niacin and marine omega-3 fatty acids (EPA andDHA) have been shown to reduce cardiovascular disease, coronary heartdisease, atherosclerosis and reduce mortality in patients withdyslipidemia, hypercholesterolemia, or Type 2 diabetes, and metabolicdisease. Niacin at high dose (1.5 to 4 grams per day) has been shown toimprove very low-density lipoprotein (“VLDL”) levels through loweringApolipoprotein B (“ApoB”) and raising high density lipoprotein (“HDL”)through increasing Apolipoprotein A1 (“ApoA1”) in the liver. Niacin canalso inhibit diacylglycerol acyltransferase-2, a key enzyme for TGsynthesis (Kamanna, V. S.; Kashyap, M. L. Am. J. Cardiol. 2008. 101(8A), 20B-26B). Unfortunately, niacin has many actions outside of theliver that detract from its therapeutic utility. the most common sideeffect of niacin is flushing, which can limit the dose a patient cantolerate. Flushing is thought to occur through the GPR 109 receptor inthe vasculature.

Omega-3 fatty acids have been shown to improve insulin sensitivity andglucose tolerance in normoglycemic men and in obese individuals. Omega-3fatty acids have also been shown to improve insulin resistance in obeseand non-obese patients with an inflammatory phenotype. Lipid, glucose,and insulin metabolism have been shown to be improved in overweighthypertensive subjects through treatment with omega-3 fatty acids.Omega-3 fatty acids (EPA/DHA) have also been shown to decreasetriglycerides and to reduce the risk for sudden death caused by cardiacarrhythmias in addition to improve mortality in patients at risk of acardiovascular event. Omega-3 fatty acids have also been taken as partof the dietary supplement portion of therapy used to treat dyslipidemia.

the ability to provide the effects of niacin and omega-3 fatty acid in asynergistic way would provide a great benefit in treating theaforementioned diseases.

SUMMARY OF THE INVENTION

This invention is based in part on the discovery of fatty acid niacinconjugates and their demonstrated effects in achieving improvedtreatment that cannot be achieved by administering niacin or fatty acidsalone or in combination. The invention is also based in part on thediscovery of niacin conjugated fatty acid mixtures and theirdemonstrated effects in achieving improved treatment that cannot beachieved by administering niacin or fatty acids alone, or incombination. These novel conjugates are useful in the treatment orprevention of metabolic diseases including atherosclerosis,dyslipidemia, coronary heart disease, hypercholesterolemia, Type 2diabetes, elevated cholesterol, metabolic syndrome and cardiovasculardisease.

Accordingly in one aspect, a molecular conjugate is described whichcomprises a niacin covalently linked to a fatty acid, wherein the fattyacid is selected from the group consisting of omega-3 fatty acids andfatty acids that are metabolized in vivo to omega-3 fatty acids, theconjugate comprises at least one amide, and the conjugate is capable ofhydrolysis to produce free niacin and free fatty acid.

In another aspect, compounds of the Formula I are described:

and pharmaceutically acceptable salts, hydrates, solvates, prodrugs,enantiomers and stereoisomers thereof;wherein,

-   -   R₁, R₂, and R₃ are each independently selected from the group        consisting of —H, -D, —Cl, —F, —CN, —NH₂, —NH(C₁-C₃ alkyl),        —N(C₁-C₃ alkyl)₂, —NH(C(O)C₁-C₃ alkyl), —N(C(O)C₁-C₃ alkyl)₂,        —C(O)H, —C(O)C₁-C₃ alkyl, —C(O)OC₁-C₃ alkyl, —C(O)NH₂,        —C(O)NH(C₁-C₃ alkyl), —C(O)N(C₁-C₃ alkyl)₂, —C₁-C₃ alkyl,        —O—C₁-C₃ alkyl, —S(O)C₁-C₃ alkyl and —S(O)₂C₁-C₃ alkyl;    -   W₁ and W₂ are each independently null O, S, NH, NR, or W₁ and W₂        can be taken together can form an imidazolidine or piperazine        group, with the proviso that W₁ and W₂ can not be O        simultaneously;    -   each a, b, c, and d is independently —H, -D, —CH₃, —OCH₃,        —OCH₂CH₃, —C(O)OR, or —O—Z, or benzyl, or two of a, b, c, and d        can be taken together, along with the single carbon to which        they are bound, to form a cycloalkyl or heterocycle;    -   each n, o, p, and q is independently 0 or 1;    -   each L is independently —O—, —S—, —S(O)—, —S(O)₂—, —S—S—,

-   -   wherein the representation of L is not limited directionally        left to right as as depicted, rather either the left side or the        right side of L can be bound to the W₁ side of the compound of        Formula I;    -   each g is independently 2, 3 or 4;    -   each h is independently 1, 2, 3 or 4;    -   m is 0, 1, 2, or 3;    -   each R₆ is independently H or C₁-C₆ alkyl, or both R₆ groups,        when taken together with the nitrogen to which they are        attached, form a heterocycle;    -   each R₇ is independently e, H, or straight or branched C₁-C₁₀        alkyl which can be optionally substituted with OH, NH₂, CO₂R,        CONH₂, phenyl, C₆H₄OH, imidazole or arginine;    -   each e is independently H or any one of the side chains of the        naturally occurring amino acids;    -   each Z is independently —H, or

-   -   with the proviso that there is at least one

-   -   in the compound;    -   each r is independently 2, 3, or 7;    -   each s is independently 3, 5, or 6;    -   each t is independently 0 or 1;    -   each v is independently 1, 2, or 6;    -   R₄ and R₅ are each independently —H, -D, —C₁-C₄ alkyl, -halogen,        —OH, —C(O)C₁-C₄ alkyl, —O-aryl, —O-benzyl, —OC(O)C₁-C₄ alkyl,        —C₁-C₃ alkene, —C₁-C₃ alkyne, —C(O)C₁-C₄ alkyl, —NH₂, —NH(C₁-C₃        alkyl, —N(C₁-C₃ alkyl)₂, —NH(C(O)C₁-C₃ alkyl), —N(C(O)C₁-C₃        alkyl)₂, —SH, —S(C₁-C₃ alkyl), —S(O)C₁-C₃ alkyl, —S(O)₂C₁-C₃        alkyl; and    -   each R is independently —H, —C(O)—C₁-C₃ alkyl, or straight or        branched C₁-C₄ alkyl optionally substituted with OR, NR₂, or        halogen;    -   provided that        -   when m, n, o, p, and q are each 0, W₁ and W₂ are each null,            and Z is

-   -   -   then t must be 0; and        -   when each of m, n, o, p, and q is 0, and W₁ and W₂ and are            each null, then Z must not be

In Formula I, any one or more of R may be substituted with a deuterium.It is also understood in Formula I that a methyl substituent can besubstituted with a C₁-C₆ alkyl.

Also described are pharmaceutical formulations comprising at least onefatty acid niacin derivative.

Also described herein are methods of treating a disease susceptible totreatment with a fatty acid niacin derivative in a patient in needthereof by administering to the patient an effective amount of a fattyacid niacin derivative.

Also described herein are methods of treating metabolic diseases byadministering to a patient in need thereof an effective amount of afatty acid niacin derivative.

The invention also includes pharmaceutical compositions that comprise aneffective amount of a fatty acid niacin derivative and apharmaceutically acceptable carrier. The compositions are useful fortreating or preventing a metabolic disease. The invention includes afatty acid niacin derivative provided as a pharmaceutically acceptableprodrug, hydrate, salt, enantiomer, stereoisomer, or mixtures thereof.

Also described herein are pharmaceutically active niacin conjugatedfatty acid mixtures which comprise a mixture of a Niacin-EPA conjugateand a Niacin-DHA conjugate. Processes for producing a niacin conjugatedfatty acid mixture are described which comprise reacting niacin, or ananalog thereof, with a mixture of EPA and DHA such that a mixture ofniacin conjugated fatty acids is produced wherein the fatty acids are amixture of DHA and EPA. In some embodiments, the mixture of DHA and EPAare esters of the fatty acids. In other embodiments, the mixture of DHAand EPA are in the free acid form. The mixture produced reflects thestarting ratios of DHA to EPA in the fatty acid mixture. For example, a50:50 mixture of EPA ester (or acid) and DHA ester (or acid) as thestarting material provides a 50:50 mixture of the Niacin-EPA conjugatesand Niacin-DHA conjugates. Similarly, a 60:40 mixture of EPA ester (oracid) and DHA ester (or acid) as the starting material provides a 60:40mixture of the Niacin-EPA conjugates and Niacin-DHA conjugates, and soon. In some instances the ratio of the mixture of EPA ester (or acid)and DHA ester (or acid) is about 60:40. In other instances, it is about40:60. The ratio of the mixture of EPA ester (or acid) and DHA ester (oracid) can be about 10:90, about 20:80, or about 30:70. In some instancesthe ratio of the mixture of EPA ester (or acid) and DHA ester (or acid)is about 90:10, about 80:20, or about 70:30. The ratio of the mixture ofEPA ester (or acid) and DHA ester (or acid) can be about 5:95, 15:85,25:75, 35:65, 45:55, 55:45, 65:35, 75:25, 85:15, or 95:5. In someinstances, the ratio of the mixture of EPA ester (or acid) and DHA ester(or acid) is between about 10:90 to about 90:10; between about 20:80 toabout 80:20; between about 25:75 to about 75:25, between about 40:60 toabout 60:40. In some instances, the ratio of the mixture of EPA ester(or acid) and DHA ester (or acid) is about 1:1. In some instances, theratio of the mixture of EPA ester (or acid) and DHA ester (or acid) isabout 1.1:1. In some instances, the ratio of the mixture of EPA ester(or acid) and DHA ester (or acid) is about 1.2:1. In some Instances, theratio of the mixture of EPA ester (or acid) and DHA ester (or acid) isabout 1.3:1. In some instances, the ratio of the mixture of EPA ester(or acid) and DHA ester (or acid) is about 1.4:1. In one instance, theratio of the mixture of EPA ester (or acid) and DHA ester (or acid) isabout 1.27:1. In another aspect the mixture of niacin conjugated fattyacids is produced by combining Niacin-EPA conjugate and Niacin-DHAtogether.

Also described herein are methods of treating a disease susceptible totreatment with a niacin conjugated fatty acid mixtures in a patient inneed thereof by administering to the patient an effective amount of aniacin conjugated fatty acid mixture.

Also described herein are methods of treating metabolic disorders byadministering to a patient in need thereof an effective amount of aniacin conjugated fatty acid mixture.

The invention also includes pharmaceutical compositions that comprise aneffective amount of a niacin conjugated fatty acid mixture and apharmaceutically acceptable carrier. The compositions are useful fortreating or preventing a metabolic disorder.

The details of the invention are set forth in the accompanyingdescription below. Although methods and materials similar or equivalentto those described herein can be used in the practice or testing of thepresent invention, illustrative methods and materials are now described.Other features, objects, and advantages of the invention will beapparent from the description and from the claims. In the specificationand the appended claims, the singular forms also include the pluralunless the context clearly dictates otherwise. Unless defined otherwise,all technical and scientific terms used herein have the same meaning ascommonly understood by one of ordinary skill in the art to which thisinvention belongs. All patents and publications cited in thisspecification are incorporated herein by reference in their entireties.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a depiction of the effect of compound I-7 on ApoB secretion inHepG2 cells.

FIG. 2 is a depiction of the effect of fatty acid niacin derivatives onSREBP-1c target genes.

DETAILED DESCRIPTION OF THE INVENTION

Metabolic diseases are a wide variety of medical disorders thatinterfere with a subject's metabolism. Metabolism is the process asubject's body uses to transform food into energy. Metabolism in asubject with a metabolic disease is disrupted in some way. The fattyacid niacin derivatives possess the ability to treat or preventmetabolic diseases.

The fatty acid niacin derivatives have been designed to bring togetherniacin analogs and omega-3 fatty acids into a single molecularconjugate. The activity of the fatty acid niacin derivatives issubstantially greater than the sum of the individual components of themolecular conjugate, suggesting that the activity induced by the fattyacid niacin derivatives is synergistic.

Definitions

The following definitions are used in connection with the fatty acidniacin derivatives:

The term “fatty acid, niacin derivatives” includes any and all possibleisomers, stereoisomers, enantiomers, diastereomers, tautomers,pharmaceutically acceptable salts, hydrates, solvates, and prodrugs ofthe fatty acid niacin derivatives described herein.

The term “niacin conjugated fatty acid mixtures” includes any and allpossible isomers, stereoisomers, enantiomers, diastereomers, tautomers,pharmaceutically acceptable salts, hydrates, solvates, and prodrugs ofthe niacin conjugated fatty acids described herein. The terms Niacin-DHAand Niacin-EPA include conjugates that are linked through bivalentlinkers between the Niacin and the DHA or EPA.

The articles “a” and “an” are used in this disclosure to refer to one ormore than one (i.e., to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

The term “and/or” is used in this disclosure to mean either “and” or“or” unless indicated otherwise.

Unless otherwise specifically defined, the term “aryl” refers to cyclic,aromatic hydrocarbon groups that have 1 to 2 aromatic rings, includingmonocyclic or bicyclic groups such as phenyl, biphenyl or naphthyl.Where containing two aromatic rings (bicyclic, etc.), the aromatic ringsof the aryl group may be joined at a single point (e.g., biphenyl), orfused (e.g., naphthyl). The aryl group may be optionally substituted byone or more substituents, e.g., 1 to 5 substituents, at any point ofattachment. The substituents can themselves be optionally substituted.

“C₁-C₃ alkyl” refers to a straight or branched chain saturatedhydrocarbon containing 1-3 carbon atoms. Examples of a C₁-C₃ alkyl groupinclude, but are not limited to, methyl, ethyl, propyl and isopropyl.

“C₁-C₄ alkyl” refers to a straight or branched chain saturatedhydrocarbon containing 1-4 carbon atoms. Examples of a C₁-C₄ alkyl groupinclude, but are not limited to, methyl, ethyl, propyl, butyl,isopropyl, isobutyl, sec-butyl and tert-butyl.

“C₁-C₅ alkyl” refers to a straight or branched chain saturatedhydrocarbon containing 1-5 carbon atoms. Examples of a C₁-C₅ alkyl groupinclude, but are not limited to, methyl, ethyl, propyl, butyl, pentyl,isopropyl, isobutyl, sec-butyl and tert-butyl, isopentyl, and neopentyl.

“C₁-C₆ alkyl” refers to a straight or branched chain saturatedhydrocarbon containing 1-6 carbon atoms. Examples of a C₁-C₆ alkyl groupinclude, but are not limited to, methyl, ethyl, propyl, butylpentyl,hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, andneopentyl.

The term, “cycloalkyl” refers to a cyclic hydrocarbon containing 3-6carbon atoms. Examples of a cycloalkyl group include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

It is understood that any of the substitutable hydrogens on an alkyl andcycloalkyl can be substituted with halogen, C₁-C₃ alkyl, hydroxyl,alkoxy and cyano groups.

The term “heterocycle” as used herein refers to a cyclic hydrocarboncontaining 3-6 atoms wherein at least one of the atoms is an O, N, or S.Examples of heterocycles include, but are not limited to, aziridine,oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine,tetrahydrofuran, tetrahydrothiophene, piperidine, tetrahydropyran,thiane, imidazolidine, oxazolidine, thiazolidine, dioxolane, dithiolane,piperazine, oxazine, dithiane, and dioxane.

The term “any one of the side chains of the naturally occurring aminoacids” as used herein means a side chain of any one of the followingamino acids: Isoleucine, Alanine, Leucine, Asparagine, Lysine,Aspartate, Methionine, Cysteine, Phenylalanine, Glutamate, Threonine,Glutamine, Tryptophan, Glycine, Valine, Proline, Arginine, Serine,Histidine, and Tyrosine.

The term “fatty acid” as used herein means an omega-3 fatty acid andfatty acids that are metabolized in vivo to omega-3 fatty acids.Non-limiting examples of fatty acids areall-cis-7,10,13-hexadecatrienoic acid, α-linolenic acid (ALA orall-cis-9,12,15-octadecatrienoic acid), stearidonic acid (STD orall-cis-6,9,12,15octadecatetraenoic acid), eicosatrienoic acid (ETE orall-cis-11,14,17-eicosatrienoic acid), eicosatetraenoic acid (ETA orall-cis-8,11,14,17-eicosatetraenoic acid), eicosapentaenoic acid (EPA orall-cis-5,8,11,14,17-eicosapentaenoic acid), docosapentaenoic acid (DPA,clupanodonic acid or all-cis-7,10,13,16,19-docosapentaenoic acid),docosahexaenoic acid (DHA or all-cis-4,7,10,13,16,19-docosahexaenoicacid), tetracosapentaenoic acid (all-cis-9,12,15,18,21-docosahexaenoicacid), or tetracosahexaenoic acid (nisinic acid orall-cis-6,9,12,15,18,21-tetracosenoic acid).

The term “niacin” as used herein means the molecule known as niacin andany derivative thereof.

A “subject” is a mammal, e.g., a human, mouse, rat, guinea pig, dog,cat, horse, cow, pig, or non-human primate, such as a monkey,chimpanzee, baboon or rhesus, and the terms “subject” and “patient” areused interchangeably herein.

The invention also includes pharmaceutical compositions comprising aneffective amount of a fatty acid niacin derivative and apharmaceutically acceptable carrier. The invention includes a fatty acidniacin derivative provided as a pharmaceutically acceptable prodrug,hydrate, salt, such as a pharmaceutically acceptable salt, enantiomers,stereoisomers, or mixtures thereof.

Representative “pharmaceutically acceptable salts” include, e.g.,water-soluble and water-insoluble salts, such as the acetate, amsonate(4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzonate,bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium,calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate,dihydrochloride, edetate, edisylate, estolate, esylate, fumarate,gluceptate, gluconate, glutamate, glycollylarsanilate,hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide,hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate,lactobionate, laurate, magnesium, malate, maleate, mandelate, mesylate,methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate,N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate,oxalate, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3-naphthoate,einbonate), pantothenate, phosphate/diphosphate, picrate,polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate,subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate,tartrate, teoclate, tosylate, triethiodide, and valerate salts.

The term “metabolic disease” as used herein refers to disorders,diseases and syndromes involving dyslipidemia, and the terms metabolicdisorder, metabolic disease, and metabolic syndrome are usedinterchangeably herein.

An “effective amount” when used in connection with a fatty acid niacinderivative is an amount effective for treating or preventing a metabolicdisease.

The term “carrier”, as used in this disclosure, encompasses carriers,excipients, and diluents and means a material, composition or vehicle,such as a liquid or solid filler, diluent, excipient, solvent orencapsulating material, involved in carrying or transporting apharmaceutical agent from one organ, or portion of the body, to anotherorgan, or portion of the body.

The term “treating”, with regard to a subject, refers to improving atleast one symptom of the subject's disorder. Treating can be curing,improving, or at least partially ameliorating the disorder.

The term “disorder” is used in this disclosure to mean, and is usedinterchangeably with, the terms disease, condition, or illness, unlessotherwise indicated.

The term “administer”, “administering”, or “administration” as used inthis disclosure refers to either directly administering a compound orpharmaceutically acceptable salt of the compound or a composition to asubject, or administering a prodrug derivative or analog of the compoundor pharmaceutically acceptable salt of the compound or composition tothe subject, which can form an equivalent amount of active compoundwithin the subject's body.

The term “prodrug,” as used in this disclosure, means a compound whichis convertible in vivo by metabolic means (e.g., by hydrolysis) to afatty acid niacin derivative.

The term “about”, as used herein, refers to ±10% of the value or rangemodified by the term “about”.

The following abbreviations are used herein and have the indicateddefinitions: Boc and BOC are tert-butoxycarbonyl, Boc₂O is di-tert-butyldicarbonate, BSA is bovine serum albumin, CDI is1,1′-carbonyldiimidazole, DCC is N,N′-dicyclohexylcarbodiimide, DIEA isN,N-diisopropylethylamine, DMAP is d-dimethylaminopyridine, DMEM isDulbecco's Modified Eagle Medium, DMF is N,N-dimethylformamide, DOSS issodium dioctyl sulfosuccinate, EDC and EDCI are1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, ELISA isenzyme-linked immunosorbent assay, EtOAc is ethyl acetate, FBS is fetalbovine serum, h is hour, HATU is2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate, HIV is human immunodeficiency virus, HPMC ishydroxypropyl methylcellulose, oxone is potassium peroxymonosulfate,Pd/C is palladium on carbon, TFA is trifluoroacetic acid, TGPS istocopherol propylene glycol succinate, and THF is tetrahydrofuran.

Compounds

Accordingly in one aspect, the present invention provides a molecularconjugate which comprises a niacin and a fatty acid covalently linked,wherein the fatty acid is selected from the group consisting of omega-3fatty acids and fatty acids that are metabolized in vivo to omega-3fatty acids, wherein the conjugate comprises at least one amide and theconjugate is capable of hydrolysis to produce free niacin and free fattyacid.

In some embodiments, the fatty acid is selected from the groupconsisting of all-cis-7,10,13-hexadecatrienoic acid, α-linolenic acid,stearidonic acid, eicosatrienoic acid, eicosatetraenoic acid,eicosapentaenoic acid (EPA), docosapentaenoic acid, docosahexaenoic acid(DHA), tetracosapentaenoic acid and tetracosahexaenoic acid. In otherembodiments, the fatty acid is selected from eicosapentaenoic acid anddocosahexaenoic acid. In some embodiments, the hydrolysis is enzymatic.

In another aspect, the present invention provides fatty acid niacinderivatives according to Formula I:

and pharmaceutically acceptable salts, hydrates, solvates, prodrugs,enantiomers, and stereoisomers thereof;wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R, W₁, W₂, L, a, c, b, d, e, g, h,m, n, o, p, q, Z, r, s, t, and v are as defined above for Formula I,

-   -   with the proviso that there is at least one

-   -   in the compound.

In some embodiments, R₃ is Cl, F, or CN.

In some embodiments, R₃ is —CH₃ or —CH₂CH₃.

In some embodiments, W₁ is NH.

In some embodiments, W₂ is NH.

In some embodiments, W₁ is O.

In some embodiments, W₂ is O.

In some embodiments, a and c are-each independently H, or CH₃.

In some embodiments, m is 0.

In other embodiments, m is 1.

In some embodiments, L is —S, or —S—S—.

In some embodiments, L is —O—.

In some embodiments, L is

In some embodiments, L is

In some embodiments, L is

In some embodiments, L is

In some embodiments, L is

In some embodiments, L is

In some embodiments, one b is O—Z, Z is

and t is 1.

In some embodiments, one d is C(O)OR.

In some embodiments n, o, p, and q are each 1.

In some embodiments, two of n, o, p, and q are each 1.

In other embodiments, three of n, o, p, and q are each 1.

In some embodiments, one Z is

and r is 2.

In some embodiments, one Z is

and r is 3.

In some embodiments, one Z is

In other embodiments, one Z is

and s is 3.

In some embodiments, one Z is

and s is 5.

In some embodiments, one Z is

and s is 6.

In some embodiments, one Z is

and v is 1.

In other embodiments, one Z is

and v is 2.

In some embodiments, one Z is

and v is 6.

In some embodiments, one Z is

and s is 3.

In some embodiments, one Z is

and s is 5.

In other embodiments, one Z is

and s is 6.

In some embodiments, t is 1.

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, n,o, p, and q are each 1, and L is O.

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, n,o, p, and q are each 1, and L is —S—S—.

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, nand o are each 0, p and q are each 1, and L is

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, n,o, p, and q are each 0, and L is

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m, n, and oare each 0, and p and q are each 1.

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, nand o are each 0, p and q are each 1, and L is

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, nand o are each 1, p and q are each 0, and L is

In some embodiments, r is 2, s is 6, m is 1, n and o are each 0, p and qare each 1, and L is

In some embodiments, r is 2, s is 6, m is 1, n and o are each 1, p and qare each 0, and L is

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, n,o, p, and q are each 1, and L is

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, n,o, p, and q are each 1, and L is NR₆.

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m, n, and oare each 0, and p and q are each 1, and one c is —CH₃ and the other c is—CH₃.

In some embodiments, r is 2, s is 6, W₁ and W₂ are each NH, m is 1, nand o are each 1, p and q are each 0, and L is

In some embodiments, r is 3, s is 5, and L is —S—S—.

In some embodiments, r is 3, s is 5, and L is —O—.

In some embodiments, r is 3, s is 5, and L is

In some embodiments, r is 3, s is 5, and L is

In some embodiments, r is 3, s is 5, and L is

In some embodiments, r is 3, s is 5, and L is

In some embodiments, r is 3, s is 5, and n, o, p, and q are each 1.

In some embodiments, r is 3, s is 5, and two of n, o, p and q are each1.

In some embodiments, r is 3, s is 5, and W₁ and W₂ are each NH.

In some embodiments, r is 3, s is 5, m is 1, n, o, p, and q are each 1,and L is O.

In some embodiments, r is 3, s is 5, m is 1, n, o, p, and q are each 1,and L is —S—S—.

In some embodiments, r is 3, s is 5, m is 1, n and o are each 0, p and qare each 1, and L is

In some embodiments, r is 3, s is 5, m is 1, n, o, p, and q are each 0,and L is

In some embodiments, r is 3, s is 5, m, n, and o are each 0, and p and qare each 1.

In some embodiments, r is 3, s is 5, m is 1, n and o are each 1, p and qare each 0, and L is

In some embodiments, r is 3, s is 5, m is 1, n and o are each 0, p and qare each 1, and L is

In some embodiments, r is 3, s is 5, m is 1, n and o are each 0, p and qare each 1, and L is

In some embodiments, r is 3, s is 5, m is 1, n and o are each 1, p and qare each 0, and L is

In some embodiments, r is 3, s is 5, m is 1, n, o, p, and q are each 1,and L is NR₆.

In some embodiments, r is 3, s is 5, m, n, and o are each 0, and p and qare each 1, and one c is —CH₃ and the other c is —CH₃.

In some embodiments, r is 3, s is 5, m is 1, n and o are each 1, p and qare each 0, and L is

In Formula I, any one or more of H may be substituted with a deuterium.It is also understood in Formula I that a methyl substituent can besubstituted with a C₁-C₆ alkyl.

In other illustrative embodiments, compounds of Formula I are as setforth below:

-   N-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethyl)nicotinamide    (I-1);-   N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(methyl)amino)ethyl)nicotinamide    (I-2);-   N-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)disulfanyl)ethyl)nicotinamide    (I-3);-   N-(2-(1-(2-(1Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)-2,5-dioxopyrrolidin-3-ylthio)ethyl)nicotinamide    (I-4);-   Methyl    3-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoacetoxy)-2-(nicotinamido)butanoate    (I-5);-   1,3-dihydroxypropan-2-yl    6-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-(nicotinamido)hexanoate    (I-6);-   N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)nicotinamide    (I-7);-   N-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide    (I-8);-   (2S,3R)-methyl    3-((S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoyloxy)-2-(nicotinamido)butanoate    (I-9);-   (2S,3R)-methyl    3-((S)-2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenamido)propanoyloxy)-2-(nicotinamido)butanoate    (I-10);-   (S)-methyl    6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)hexanoate    (I-11);-   (S)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)hexanoic    acid (I-12);-   (S)-methyl    2-((5Z,8Z,1Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-6-(nicotinamido)hexanoate    (I-13);-   (S)-2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-6-(nicotinamido)hexanoic    acid (I-14);-   (S)-methyl    6-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-2-(nicotinamido)hexanoate    (I-15);-   (S)-6-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-2-(nicotinamido)hexanoic    acid (I-16);-   (S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-6-(nicotinamido)hexanoic    acid (I-17);-   (S)-5-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)pentanoic    acid (I-18);-   (S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-5-(nicotinamido)pentanoic    acid (I-19);-   4-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-(nicotinamido)butanoic    acid (I-20);-   2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-4-(nicotinamido)butanoic    acid (I-21);-   3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-(nicotinamido)propanoic    acid (I-22);-   2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-3-(nicotinamido)propanoic    acid (I-23);-   2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)-4-(nicotinamido)butanoic    acid (I-24);-   (S)-1,3-dihydroxypropan-2-yl    2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-6-(nicotinamido)hexanoate    (I-25);-   (S)-1,3-dihydroxypropan-2-yl    5-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)pentanoate    (I-26);-   (S)-1,3-dihydroxypropan-2-yl    2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-5-(nicotinamido)pentanoate    (I-27);-   1,3-dihydroxypropan-2-yl    4-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-(nicotinamido)butanoate    (I-28);-   1,3-dihydroxypropan-2-yl    2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-4-(nicotinamido)butanoate    (I-29);-   1,3-dihydroxypropan-2-yl    3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-(nicotinamido)propanoate    (I-30);-   1,3-dihydroxypropan-2-yl    2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-3-(nicotinamido)propanoate    (I-31);-   1,3-dihydroxypropan-2-yl    2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)-4-(nicotinamido)butanoate    (I-32);-   N-(4-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidobutyl)nicotinamide    (I-33);-   N-(3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidopropyl)nicotinamide    (I-34);-   N-(1-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-methylpropan-2-yl)nicotinamide    (I-35);-   N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-methylpropyl)nicotinamide    (I-36);-   N-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethylamino)ethyl)nicotinamide    (I-37);-   N-(3-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethylamino)propyl)nicotinamide    (I-38);-   N-(2-(3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidopropylamino)ethyl)nicotinamide    (I-39);-   N-(2-((3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidopropyl)(ethyl)amino)ethyl)nicotinamide    (I-40);-   N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)isobutyl)amino)ethyl)nicotinamide    (I-41);-   N-(2-(N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)acetamido)ethyl)nicotinamide    (I-42);-   N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(2-morpholinoethyl)amino)ethyl)nicotinamide    (I-43);-   N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(3-(piperazin-1-yl)propyl)amino)ethyl)nicotinamide    (I-44);-   N-(3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-oxopropyl)nicotinamide    (I-45);-   N-(3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-morpholinopropyl)nicotinamide    (I-46);-   N-(3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-2-(piperazin-1-yl)propyl)nicotinamide    (I-47);-   N-(3-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-4-(4-methylpiperazin-1-yl)propyl)nicotinamide    (I-48);-   N-(5-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-3-hydroxypentyl)nicotinamide    (I-49);-   N-(5-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-3-morpholinopentyl)nicotinamide    (I-50);-   N-(5-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido-3-(piperazin-1-yl)pentyl)nicotinamide    (I-51);-   (S)-((R)-1-(nicotinamido)propan-2-yl)    2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)propanoate    (I-52);-   (S)-((R)-1-(nicotinamido)propan-2-yl)    2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-3-methylbutanoate    (I-53);-   N-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethoxy)ethyl)nicotinamide    (I-54);-   N-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)docosa-4,7,10,13,16,19-hexaenamidoethylthio)ethyl)nicotinamide    (I-55);-   (4Z,7Z,10Z,13Z,16Z,19Z)-1-(nicotinamido)propan-2-yl    docosa-4,7,10,13,16,19-hexaenoate (I-56);-   (4Z,7Z,10Z,13Z,16Z,19Z)-4-methoxy-3-(nicotinamido)-4-oxobutan-2-yl    docosa-4,7,10,13,16,19-hexaenoate (I-57);-   N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)-6-methylnicotinamide    (I-58);-   N-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)-6-methylnicotinamide    (I-59);-   N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)-6-ethylnicotinamide    (I-60);-   6-ethyl-N-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide    (I-61);-   6-chloro-N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)nicotinamide    (I-62);-   6-chloro-N-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide    (I-63);-   N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)-6-fluoronicotinamide    (I-64);-   6-fluoro-N-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide    (I-65);-   6-cyano-N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)nicotinamide    (I-66);-   6-cyano-N-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide    (I-67);-   (S)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(2-methylnicotinamido)hexanoic    acid (I-68);-   (S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-6-(2-methylnicotinamido)hexanoic    acid (I-69);-   (S)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(2-ethylnicotinamido)hexanoic    acid (I-70);-   (S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-6-(2-ethylnicotinamido)hexanoic    acid (I-71);-   (S)-2-(2-chloronicotinamido)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)hexanoic    acid (I-72);-   (S)-6-(2-chloronicotinamido)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)hexanoic    acid (I-73);-   (S)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(2-fluoronicotinamido)hexanoic    acid (I-74);-   (S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-6-(2-fluoronicotinamido)hexanoic    acid (I-75);-   (S)-2-(2-cyanonicotinamido)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)hexanoic    acid (I-76);-   (S)-6-(2-cyanonicotinamido)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)hexanoic    acid (I-77);-   N-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethyl)-6-methylnicotinamide    (I-78);-   N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(methyl)amino)ethyl)-6-methylnicotinamide    (I-79);-   N-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)disulfanyl)ethyl)-6-methylnicotinamide    (I-80);-   N-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethyl)-6-ethylnicotinamide    (I-81);-   N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(methyl)amino)ethyl)-6-ethylnicotinamide    (I-82);-   N-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)disulfanyl)ethyl)-6-ethylnicotinamide    (I-83);-   6-chloro-N-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethyl)nicotinamide    (I-84);-   6-chloro-N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(methyl)amino)ethyl)nicotinamide    (I-85);-   6-chloro-N-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)disulfanyl)ethyl)nicotinamide    (I-86);-   6-cyano-N-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethyl)nicotinamide    (I-87);-   6-cyano-N-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(methyl)amino)ethyl)nicotinamide    (I-88);-   6-cyano-N-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)disulfanyl)ethyl)nicotinamide    (I-89);

Mixtures

The present invention also provides mixtures of a Niacin-EPA conjugateand a Niacin-DHA conjugate. The mixtures can contain Niacin-EPAconjugates or Niacin-DHA conjugates in any proportion. The ratio ofNiacin-EPA conjugate to the Niacin-DHA conjugate reflects the ratio ofthe EPA ester (or acid) to the DHA ester (or acid) present in thestarting mixture of DHA and EPA esters (or acids). Also described hereinare pharmaceutically active niacin conjugated fatty acid mixtures whichcomprise a mixture of a Niacin-EPA conjugate and a Niacin-DHA conjugate.Processes for producing a niacin conjugated fatty acid mixture aredescribed which comprise reacting niacin, or an analog thereof, with amixture of EPA and DHA such that a mixture of niacin conjugated fattyacids is produced wherein the fatty acids are a mixture of DHA and EPA.In some embodiments, the mixture of DHA and EPA are esters of the fattyacids. In other embodiments, the mixture of DHA and EPA are in the freeacid form. The mixture produced reflects the starting ratios of DHA toEPA in the fatty acid mixture. For example, a 50:50 mixture of EPA ester(or acid) and DHA ester (or acid) as the starting material provides a50:50 mixture of the Niacin-EPA conjugates and Niacin-DHA conjugates.Similarly, a 60:40 mixture of EPA ester (or acid) and DHA ester (oracid) as the starting material provides a 60:40 mixture of theNiacin-EPA conjugates and Niacin-DHA conjugates, and so on. In someinstances the ratio of the mixture of EPA ester (or acid) and DHA ester(or acid) is about 60:40. In other instances, it is about 40:60. Theratio of the mixture of EPA ester (or acid) and DHA ester (or acid) canbe about 10:90, about 20:80, or about 30:70. In some instances the ratioof the mixture of EPA ester (or acid) and DHA ester (or acid) is about90:10, about 80:20, or about 70:30. The ratio of the mixture of EPAester (or acid) and DHA ester (or acid) can be about 5:95, 15:85, 25:75,35:65, 45:55, 55:45, 65:35, 75:25, 85:15, or 95:5. In some instances,the ratio of the mixture of EPA ester (or acid) and DHA ester (or acid)is between about 10:90 to about 90:10; between about 20:80 to about80:20; between about 25:75 to about 75:25, between about 40:60 to about60:40. In some instances, the ratio of the mixture of EPA ester (oracid) and DHA ester (or acid) is about 1:1. In some instances, the ratioof the mixture of EPA ester (or acid) and DHA ester (or acid) is about1.2:1. In some instances, the ratio of the mixture of EPA ester (oracid) and DHA ester (or acid) is about 1.3:1. In some instances, theratio of the mixture of EPA ester (or acid) and DHA ester (or acid) isabout 1.4:1. In one instance, the ratio of the mixture of EPA ester (oracid) and DHA ester (or acid) is about 1.27:1. In another aspect themixture of niacin conjugated fatty acids is produced by combiningNiacin-EPA conjugate and Niacin-DHA together.

Methods for Using Fatty Acid Niacin Derivatives

The invention also includes methods for treating metabolic diseases suchas the treatment or prevention of metabolic diseases includingatherosclerosis, dyslipidemia, coronary heart disease,hypercholesterolemia, Type 2 diabetes, elevated cholesterol, metabolicsyndrome and cardiovascular disease.

In one embodiment, the method comprises contacting a cell with a fattyacid niacin derivative in an amount sufficient to decrease the releaseof triglycerides or VLDL or LDL or cause an increase in reversecholesterol transport or increase HDL concentrations.

Also provided in the invention is a method for inhibiting, preventing,or treating a metabolic disease, or symptoms of a metabolic disease, ina subject. Examples of such disorders include, but are not limited toatherosclerosis, dyslipidemia, hypertriglyceridemia, hypertension, heartfailure, cardiac arrhythmias, low HDL levels, high LDL levels, suddendeath, stable angina, coronary heart disease, acute myocardialinfarction, secondary prevention of myocardial infarction,cardiomyopathy, endocarditis, type 2 diabetes, insulin resistance,impaired glucose tolerance, hypercholesterolemia, stroke,hyperlipidemia, hyperlipoproteinemia, chronic kidney disease,intermittent claudication, hyperphosphatemia, carotid atherosclerosis,peripheral arterial disease, diabetic nephropathy, hypercholesterolemiain HIV infection, acute coronary syndrome (ACS), non-alcoholic fattyliver disease, arterial occlusive diseases, cerebral arteriosclerosis,cerebrovascular disorders, myocardial ischemia, and diabetic autonomicneuropathy.

In some embodiments, the subject is administered an effective amount ofa fatty acid niacin derivative.

The invention also includes pharmaceutical compositions useful fortreating or preventing a metabolic disease, or for inhibiting ametabolic disease, or more than one of these activities. Thecompositions can be suitable for internal use and comprise an effectiveamount of a fatty acid niacin derivative and a pharmaceuticallyacceptable carrier. The fatty acid niacin derivatives are especiallyuseful in that they demonstrate very low peripheral toxicity or noperipheral toxicity. In some embodiments, the pharmaceutical compositioncomprises one fatty acid niacin derivative. In other embodiments, thepharmaceutical composition comprises two fatty acid niacin derivatives.

The fatty acid niacin derivatives can each be administered in amountsthat are sufficient to treat or prevent a metabolic disease or preventthe development thereof in subjects.

Administration of the fatty acid niacin derivatives can be accomplishedvia any mode of administration for therapeutic agents. These modesinclude systemic or local administration such as oral, nasal,parenteral, transdermal, subcutaneous, vaginal, buccal, rectal ortopical administration modes.

Depending on the intended mode of administration, the compositions canbe in solid, semi-solid or liquid dosage form, such as, for example,injectables, tablets, suppositories, pills, time-release capsules,elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, orthe like, sometimes in unit dosages and consistent with conventionalpharmaceutical practices. Likewise, they can also be administered inintravenous (both bolus and infusion), intraperitoneal, subcutaneous orintramuscular form, all using forms well known to those skilled in thepharmaceutical arts.

Illustrative pharmaceutical compositions are tablets and gelatincapsules comprising a fatty acid niacin derivative and apharmaceutically acceptable carrier, such as: a) a diluent, e.g.,purified water, triglyceride oils, such as hydrogenated or partiallyhydrogenated vegetable oil, or mixtures thereof, corn oil, olive oil,sunflower oil, safflower oil, fish oils, such as EPA or DHA, or theiresters or triglycerides or mixtures thereof, omega-3 fatty acids orderivatives thereof, lactose, dextrose, sucrose, mannitol, sorbitol,cellulose, sodium, saccharin, glucose and/or glycine; b) a lubricant,e.g., silica, talcum, stearic acid, its magnesium or calcium salt,sodium oleate, sodium stearate, magnesium stearate, sodium benzoate,sodium acetate, sodium chloride and/or polyethylene glycol; for tabletsalso; c) a binder, e.g., magnesium aluminum silicate, starch paste,gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose,magnesium carbonate, natural sugars such as glucose or beta-lactose,corn sweeteners, natural and synthetic gums such as acacia, tragacanthor sodium alginate, waxes and/or polyvinylpyrrolidone, if desired; d) adisintegrant, e.g., starches, agar, methyl cellulose, bentonite, xanthangum, alginic acid or its sodium salt, or effervescent mixtures; e)absorbent, colorant, flavorant and sweetener; f) an emulsifier ordispersing agent, such as Tween 80, Labrasol, HPMC, DOSS, caproyl 909,labrafac, labrafil, peceol, transcutol, capmul MCM, capmul PG-12, captex355, gelucire, vitamin E TGPS, or other acceptable emulsifier; and/or g)an agent that enhances absorption of the compound such as cyclodextrin,hydroxypropyl-cyclodextrin, PEG-400, PEG200.

Liquid, particularly injectable, compositions can, for example, beprepared by dissolution, dispersion, etc. For example, the fatty acidniacin derivative is dissolved in or mixed with a pharmaceuticallyacceptable solvent such as, for example, water, saline, aqueousdextrose, glycerol, ethanol, and the like, to thereby form an injectableisotonic solution or suspension. Proteins such as albumin, chylomicronparticles, or serum proteins can be used to solubilize the fatty acidniacin derivatives.

The fatty acid niacin derivatives can be also formulated as asuppository that can be prepared from fatty emulsions or suspensions;using polyalkylene glycols such as propylene glycol, as the carrier.

The fatty acid niacin derivatives can also be administered in the formof liposome delivery systems, such as small unilamellar vesicles, largeunilamellar vesicles and multilamellar vesicles. Liposomes can be formedfrom a variety of phospholipids, containing cholesterol, stearylamine orphosphatidylcholines. In some embodiments, a film of lipid components ishydrated with an aqueous solution of drug to a form lipid layerencapsulating the drug, as described in U.S. Pat. No. 5,262,564, thecontents of which are herein incorporated by reference in theirentirety.

Fatty acid niacin derivatives can also be delivered by the use ofmonoclonal antibodies as individual carriers to which the fatty acidniacin derivatives are coupled. The fatty acid niacin derivatives canalso be coupled with soluble polymers as targetable drug carriers. Suchpolymers can include polyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamide-phenol,polyhydroxyethylaspanamidephenol, or polyethyleneoxidepolylysinesubstituted with palmitoyl residues. Furthermore, the fatty acid niacinderivatives can be coupled to a class of biodegradable polymers usefulin achieving controlled release of a drug, for example, polylactic acid,polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters,polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked oramphipathic block copolymers of hydrogels. In one embodiment, fatty acidniacin derivatives are not covalently bound to a polymer, e.g., apolycarboxylic acid polymer, or a polyacrylate.

Parenteral injectable administration is generally used for subcutaneous,intramuscular or intravenous injections and infusions. Injectables canbe prepared in conventional forms, either as liquid solutions orsuspensions or solid forms suitable for dissolving in liquid prior toinjection.

Compositions can be prepared according to conventional mixing,granulating or coating methods, respectively, and the presentpharmaceutical compositions can contain from about 0.1% to about 80%,from about 5% to about 60%, or from about 1% to about 20% of the fattyacid niacin derivative by weight or volume.

The dosage regimen utilizing the fatty acid niacin derivative isselected in accordance with a variety of factors including type,species, age, weight, sex and medical condition of the patient; theseverity of the condition to be treated; the route of administration;the renal or hepatic function of the patient; and the particular fattyacid niacin derivative employed. A physician or veterinarian of ordinaryskill in the art can readily determine and prescribe the effectiveamount of the drug required to prevent, counter or arrest the progressof the condition.

Effective dosage amounts of the present invention, when used for theindicated effects, range from about 20 mg to about 5,000 mg of the fattyacid niacin derivative per day. Compositions for in vivo or in vitro usecan contain about 20, 50, 75, 100, 150, 250, 500, 750, 1,000, 1,250,2,500, 3,500, or 5,000 mg of the fatty acid niacin derivative. In oneembodiment, the compositions are in the form of a tablet that can bescored. Effective plasma levels of the fatty acid niacin derivative canrange from about 0.002 mg to about 100 mg per kg of body weight per day.Appropriate dosages of the fatty acid niacin derivatives can bedetermined as set forth in Goodman, L. S.; Gilman, A. ThePharmacological Basis of Therapeutics, 5th ed.; MacMillan; New York,1975, pp. 201-226.

Fatty acid niacin derivatives can be administered in a single dailydose, or the total daily dosage can be administered in divided doses oftwo, three or four times daily. Furthermore, fatty acid niacinderivatives can be administered in intranasal form via topical use ofsuitable intranasal vehicles, or via transdermal routes, using thoseforms of transdermal skin patches well known to those of ordinary skillin that art. To be administered in the form of a transdermal deliverysystem, the dosage administration can be continuous rather thanintermittent throughout the dosage regimen. Other illustrative topicalpreparations include creams, ointments, lotions, aerosol sprays andgels, wherein the concentration of the fatty acid niacin derivativeranges from about 0.1% to about 15%, w/w or w/v.

Methods for Using Niacin Conjugated Fatty Acid Mixtures

the invention also includes methods for treating metabolic diseases suchas the treatment or prevention of metabolic diseases includingatherosclerosis, dyslipidemia, coronary heart disease,hypercholesterolemia, Type 2 diabetes, elevated cholesterol, metabolicsyndrome and cardiovascular disease.

In one embodiment, the method comprises contacting a cell with a niacinconjugated fatty acid mixture in an amount sufficient to decrease therelease of triglycerides or VLDL or LDL or cause an increase in reversecholesterol transport or increase HDL concentrations.

Also provided in the invention is a method for inhibiting, preventing,or treating a metabolic disease, or symptoms of a metabolic disease, ina subject. Examples of such disorders include, but are not limited toatherosclerosis, dyslipidemia, hypertriglyceridemia, hypertension, heartfailure, cardiac arrhythmias, low HDL levels, high LDL levels, suddendeath, stable angina, coronary heart disease, acute myocardialinfarction, secondary prevention of myocardial infarction,cardiomyopathy, endocarditis, type 2 diabetes, insulin resistance,impaired glucose tolerance, hypercholesterolemia, stroke,hyperlipidemia, hyperlipoproteinemia, chronic kidney disease,intermittent claudication, hyperphosphatemia, carotid atherosclerosis,peripheral arterial disease, diabetic nephropathy, hypercholesterolemiain HIV infection, acute coronary syndrome (ACS), non-alcoholic fattyliver disease, arterial occlusive diseases, cerebral arteriosclerosis,cerebrovascular disorders, myocardial ischemia, and diabetic autonomicneuropathy.

In some embodiments, the subject is administered an effective amount ofa niacin conjugated fatty acid mixture.

The invention also includes pharmaceutical compositions useful fortreating or preventing a metabolic disease, or for inhibiting ametabolic disease, or more than one of these activities. Thecompositions can be suitable for internal use and comprise an effectiveamount of a niacin conjugated fatty acid mixture and a pharmaceuticallyacceptable carrier. The niacin conjugated fatty acid mixtures areespecially useful in that they demonstrate very low peripheral toxicityor no peripheral toxicity.

The niacin conjugated fatty acid mixtures can each be administered inamounts that are sufficient to treat or prevent a metabolic disease orprevent the development thereof in subjects.

Administration of the niacin conjugated fatty acid mixtures can beaccomplished via any mode of administration for therapeutic agents.These modes include systemic or local administration such as oral,nasal, parenteral, transdermal, subcutaneous, vaginal, buccal, rectal ortopical administration modes.

Depending on the intended mode of administration, the compositions canbe in solid, semi-solid or liquid dosage form, such as, for example,injectables, tablets, suppositories, pills, time-release capsules,elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, orthe like, sometimes in unit dosages and consistent with conventionalpharmaceutical practices. Likewise, they can also be administered inintravenous (both bolus and infusion), intraperitoneal, subcutaneous orintramuscular form, all using forms well known to those skilled in thepharmaceutical arts.

Illustrative pharmaceutical compositions are tablets and gelatincapsules comprising a niacin conjugated fatty acid mixture and apharmaceutically acceptable carrier, such as: a) a diluent, e.g.,purified water, triglyceride oils, such as hydrogenated or partiallyhydrogenated vegetable oil, or mixtures thereof, corn oil, olive oil,sunflower oil, safflower oil, fish oils, such as EPA or DHA, or theiresters or triglycerides or mixtures thereof, omega-3 fatty acids orderivatives thereof, lactose, dextrose, sucrose, mannitol, sorbitol,cellulose, sodium, saccharin, glucose and/or glycine; b) a lubricant,e.g., silica, talcum, stearic acid, its magnesium or calcium salt,sodium oleate, sodium stearate, magnesium, stearate, sodium benzoate,sodium acetate, sodium chloride and/or polyethylene glycol; for tabletsalso; c) a binder, e.g., magnesium aluminum silicate, starch paste,gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose,magnesium carbonate, natural sugars such as glucose or beta-lactose,corn sweeteners, natural and synthetic gums such as acacia, tragacanthor sodium alginate, waxes and/or polyvinylpyrrolidone, if desired; d) adisintegrant, e.g., starches, agar, methyl cellulose, bentonite, xanthangum, alginic acid, or its sodium salt, or effervescent mixtures; e)absorbent, colorant, flavorant and sweetener; f) an emulsifier ordispersing agent, such as Tween 80, Labrasol, HPMC, DOSS, caproyl 909,labrafac, labrafil, peceol, transcutol, capmul MCM, capmul PG-12, captex355, gelucire, vitamin E TGPS or other acceptable emulsifier; and/or g)an agent that enhances absorption of the compound such as cyclodextrin,hydroxypropyl-cyclodextrin, PEG400, PEG200.

Liquid, particularly injectable, compositions can, for example, beprepared by dissolution, dispersion, etc. For example, the niacinconjugated fatty acid mixture is dissolved in or mixed with apharmaceutically acceptable solvent such as, for example, water, saline,aqueous dextrose, glycerol, ethanol and the like, to thereby form aninjectable isotonic solution or suspension. Proteins such as albumin,chylomicron particles, or serum proteins can be used to solubilize thefatty acid niacin derivatives.

The niacin conjugated fatty acid mixture can be also formulated as asuppository that can be prepared from fatty emulsions or suspensions;using polyalkylene glycols such as propylene glycol, as the carrier.

The niacin conjugated fatty acid mixture can also be administered in theform of liposome delivery systems, such as small unilamellar vesicles,large unilamellar vesicles and multilamellar vesicles. Liposomes can beformed from a variety of phospholipids, containing cholesterol,stearylamine or phosphatidylcholines. In some embodiments, a film oflipid components is hydrated with an aqueous solution of drug to a formlipid layer encapsulating the drug, as described in U.S. Pat. No.5,262,564, the contents of which are herein incorporated by reference intheir entirety.

Niacin conjugated fatty acid mixtures can also be delivered by the useof monoclonal antibodies as individual carriers to which the fatty acidniacin derivatives are coupled. The niacin conjugated fatty acidmixtures can also be coupled with soluble polymers as targetable drugcarriers. Such polymers can include polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropylmethacrylamide-phenol,polyhydroxyethylaspanamidephenol, or polyethyleneoxidepolylysinesubstituted with palmitoyl residues. Furthermore, the niacin conjugatedfatty acid mixtures can be coupled to a class of biodegradable polymersuseful in achieving controlled release of a drug, for example,polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates andcross-linked or amphipathic block copolymers of hydrogels. In oneembodiment, niacin conjugated fatty acid mixtures are not covalentlybound to a polymer, e.g., a polycarboxylic acid polymer, or apolyacrylate.

Parenteral injectable administration is generally used for subcutaneous,intramuscular or intravenous injections and infusions. Injectables canbe prepared in conventional forms, either as liquid solutions orsuspensions or solid forms suitable for dissolving in liquid prior toinjection.

Compositions can be prepared according to conventional mixing,granulating or coating methods, respectively, and the presentpharmaceutical compositions can contain from about 0.1% to about 80%,from about 5% to about 60%, or from about 1% to about 20% of the niacinconjugated fatty acid mixtures by weight or volume.

The dosage regimen utilizing the niacin conjugated fatty acid mixturesis selected in accordance with a variety of factors including type,species, age, weight, sex and medical condition of the patient; theseverity of the condition to be treated; the route of administration;the renal or hepatic function of the patient; and the particular niacinconjugated fatty acid mixture employed. A physician or veterinarian ofordinary skill in the art can readily determine and prescribe theeffective amount of the drug required to prevent, counter or arrest theprogress of the condition.

Effective dosage amounts of the present invention, when used for theindicated effects, range from about 20 mg to about 5,000 mg of theniacin conjugated fatty acid mixture per day. Compositions for in vivoor in vitro use can contain about 20, 50, 75, 100, 150, 250, 500, 750,1,000, 1,250, 2,500, 3,500, or 5,000 mg of the niacin conjugated fattyacid mixtures. In one embodiment, the compositions are in the form of atablet that can be scored. Effective plasma levels of the niacinconjugated fatty acid mixtures can range from about 0.002 mg to about100 mg per kg of body weight per day. Appropriate dosages of the niacinconjugated fatty acid mixtures can be determined as set forth inGoodman, L. S.; Gilman, A. The Pharmacological Basis of Therapeutics,5th ed.; MacMillan: New York, 1975, pp. 201-226.

Niacin conjugated fatty acid mixtures can be administered in a singledaily dose, or the total daily dosage can be administered in divideddoses of two, three or four times daily. Furthermore, niacin conjugatedfatty acid mixtures can be administered in intranasal form via topicaluse of suitable intranasal vehicles, or via transdermal routes, usingthose forms of transdermal skin patches well known to those of ordinaryskill in the art. To be administered in the form of a transdermaldelivery system, the dosage administration can be continuous rather thanintermittent throughout the dosage regimen. Other illustrative topicalpreparations include creams, ointments, lotions, aerosol sprays andgels, wherein the concentration of the niacin conjugated fatty acidmixture ranges from about 0.1% to about 15%, w/w or w/v.

METHODS OF MAKING Methods for Making the Fatty Acid Niacin Derivatives

Examples of synthetic pathways useful for making fatty acid niacinderivatives of Formula I are set forth in the Examples below andgeneralized in Schemes 1-10.

wherein R₆, r, and s are as defined above.

The mono-BOC protected amine of the formula B can be obtained fromcommercial sources or prepared according to the procedures outlined inKrapcho et al. Synthetic Communications 1990, 20, 2559-2564. Compound Acan be amidated with the amine B using a coupling reagent such as DCC,CDI, EDC, or optionally with a tertiary amine base and/or catalyst,e.g., DMAP, followed by deprotection of the BOC group with acids such asTFA or HCl in a solvent such as CH₂Cl₂ dioxane to produce the coupledcompound C. Activation of compound C with a coupling agent such as HATUin the presence of an amine such as DIEA followed by addition of a fattyacid of formula D affords compounds of the formula E.

wherein R, r, and s are as defined above.

The acylated amine of the formula F can be prepared using the proceduresoutlined in Andruszkiewicz et al. Synthetic Communications 2008, 38,905-913. Compound A can be amidated with the amine F using a couplingreagent such as DCC, CDI, EDC, or optionally with a tertiary amine baseand/or catalyst, e.g., DMAP, followed by deprotection of the BOC groupwith acids such as TFA or HCl in a solvent such as CH₂Cl₂ or dioxane toproduce the coupled compound G. Activation of compound G with a couplingagent such as HATU in the presence of an amine such as DIEA followed byaddition of a fatty acid of formula D affords compounds of the formulaH.

wherein r and s are as defined above.

Compound A can be amidated with the corresponding amine I (where i=0, 1,2 or 3) using a coupling reagent such as DCC, CDI, EDC, or optionallywith a tertiary amine base and/or catalyst, e.g., DMAP, followed bydeprotection of the BOC group with acids such as TFA or HCl in a solventsuch as CH₂Cl₂ or dioxane to produce the coupled compound J. Activationof compound J with a coupling agent such as HATU in the presence of anamine such as DIEA followed by addition of a fatty acid of formula Baffords compounds of the formula K. Hydrolysis of the ester under basicconditions such as NaOH or LiOH produces the corresponding acid, whichcan be coupled with glycidol to afford compounds of the formula L.

wherein r and s are as defined above.

The amine M can be prepared according to the procedures outlined inDahan et al. J. Org. Chem. 2007, 72, 2289-2296. Compound A can becoupled with the amine M using a coupling reagent such as DCC, CDI, EDC,or optionally with a tertiary amine base and/or catalyst, e.g., DMAP,followed by deprotection of the BOC group with acids such as TFA or HClin a solvent such as CH₂Cl₂ or dioxane to produce the coupled compoundN. Activation of compound N with a coupling agent such as HATU in thepresence of an amine such as DIEA hallowed by addition of a fatty acidof formula D affords compounds of the formula O.

wherein r and s are as defined above.

Compound A can be amidated with the commercially available amine P usinga coupling reagent such as DCC, CDI, EDC, or optionally with a tertiaryamine base and/or catalyst, e.g., DMAP, to afford compound Q. The BOCgroup in compound Q can be removed with acids such as TFA or HCl in asolvent such as CH₂Cl₂ or dioxane and the resulting amine can be coupledwith a fatty acid of formula D using a coupling agent such as HATU inthe presence of an amine such as DIEA to afford compounds of the formulaR. To those skilled in the art, the sulfur group in formula Q can beoxidized to the corresponding sulfoxide or sulfone using an oxidizingagent such as H₂O₂ or oxone.

wherein R₆, r, and s are as defined above.

The amine T can be prepared from the commercially available diamineaccording to the procedures outlined in Dahan et al. J. Org. Chem. 2007,72, 2289-2296. Compound A can be amidated with the amine T using acoupling reagent such as DCC, CDI, EDC, or optionally with a tertiaryamine base and/or catalyst, e.g., DMAP, to afford compound U. The BOCgroup of compound U can be removed with acids such as TFA or HCl in asolvent such as CH₂Cl₂ or dioxane and the resulting amine can be coupledwith a fatty acid of formula D using HATU in the presence of an aminesuch as DIEA to afford compounds of the formula V. To those skilled inthe art, the hydroxyl group in compound U can be further acylated orconverted to an amino group by standard mesylation chemistry followed bydisplacement with sodium azide and hydrogenation over a catalyst such asPd/C. The amine can be further acylated or alkylated, followed by theremoval of the BOC group. The resulting amine can be coupled with afatty acid of the formula D to afford compounds of the formula W.

wherein r and s are as defined above.

Compound A can be amidated with the commercially available amine X usinga coupling reagent such as DCC, CDI, EDC, optionally with a tertiaryamine base and/or catalyst, e.g., DMAP to afford compound Y. The BOCgroup in compound Y can be removed with acids such as TFA or HCl in asolvent such as CH₂Cl₂ or dioxane. The resulting amine can be coupledwith a fatty acid of the formula D using a coupling agent such as HATUin the presence of an amine such as DIEA to afford compounds of theformula Z.

wherein r and s are as defined above.

Compound A can be amidated with the commercially available cysteinemethyl ester using a coupling reagent such as DCC, CDI, EDC, oroptionally with a tertiary amine base and/or catalyst, e.g., DMAP, toafford compound AA. The commercially available maleimide derivative BBcan be coupled with a fatty acid of the formula D using a coupling agentsuch as HATU or EDCI to afford compounds of the formula CC. Compound AAcan be coupled to compounds of the formula CC in a solvent such asacetonitrile to afford compounds of the formula DD.

wherein R₇, a, r, and s are as defined above.

The commercially available amino acid esters EE can be coupled with afatty acid of the formula D using a coupling agent such as EDCI or HATU,followed by alkaline hydrolysis of the methyl ester to afford compoundsof the formula FF. Compounds of the formula FF can be coupled with thecommercially available BOC-amino acid derivatives GG using a couplingagent such as EDCI or HATU. The BOC group can be removed by treatmentwith acids such as TFA or HCl to afford compounds of the formula HHwhich can then be coupled with compound A to afford compounds of theformula II.

The invention also includes methods for making the niacin conjugatedfatty acid mixtures.

Mixture JJ can be hydrolyzed under basic conditions such as NaOH inaqueous THF or MeOH to produce the corresponding mixture of free fattyacids KK. Activation of mixture KK with a coupling agent such as DCC,CDI, EDC, HATU or optionally with a tertiary amine base and/or catalyst,e.g., DMAP or HOBT followed by addition of an amine of formula LLaffords niacin conjugated fatty acid mixtures of the formula MM.Alternatively, the synthesis can utilize the mixture of free fatty acidsas the starting material to produce the product MM. The amine LL can beprepared as described in Schemes 1-9 and in U.S. Ser. No. 12/872,555.

Methods for Making Niacin Conjugated Fatty Acid Mixtures

The invention also includes methods for making the niacin conjugatedfatty acid mixtures.

Mixture A can be hydrolyzed under basic conditions such as NaOH inaqueous THF or MeOH to produce the corresponding mixture of free fattyacids B. Activation of mixture B with a coupling agent such as DCC, CDI,EDC, HATU or optionally with a tertiary amine base and/or catalyst,e.g., DMAP or HOBT followed by addition of an amine of formula C affordsniacin conjugated fatty acid mixtures of the formula D. Alternatively,the synthesis can start with the mixture office fatty acids B.

EXAMPLES

The disclosure is further illustrated by the following examples, whichare not to be construed as limiting this disclosure in scope or spiritto the specific procedures herein described. It is to be understood thatthe examples are provided to illustrate certain embodiments and that nolimitation to the scope of the disclosure is intended thereby. It is tobe further understood that resort may be had to various otherembodiments, modifications, and equivalents thereof which may suggestthemselves to those skilled in the art without departing from the spiritof the present disclosure and/or scope of the appended claims.

Example 1 Effect of Fatty Acid Niacin Derivatives on ApoB Secretion inHepG2 Cells

Niacin has been reported to increase serum levels of HDL to LDLcholesterol in vivo. Similarly, niacin has been reported to increase thesecretion of ApoA1 (Jin, F-Y. et al. Arterioscler. Thromb. Vasc. Biol.1997, 17 (10), 2020-2028) while inhibiting the secretion of ApoB (Jin,F-Y. et al. Arterioscler. Thromb. Vasc. Biol. 1999, 19, 1051-1059) inthe media supernatants of HepG2 cultures. Independently, DHA has beendemonstrated to lower ApoB as well (Pan, M. et al. J. Clin. Invest.2004, 113, 1277-1287) by a very different mechanism. Thus, the secretionof ApoB from HepG2 cells possesses utility as a cell based read-out forniacin-DHA conjugates as well as mixtures thereof, as well asderivatives of same.

HepG2 cells (ATCC) are seeded at 10,000 cells per well in 96 wellplates. After adhering overnight, growth media (10% FBS in DMEM) isremoved and cells are serum starved for 24 hours in DMEM containing 0.1%fatty acid free bovine serum albumin (BSA, Sigma). Cells are thentreated with a compound. Niacin at 5 mM is used as a positive control.All treatments are performed in triplicate. Simultaneous with compoundtreatment, ApoB secretion is stimulated with addition of 0.1 oleatecomplexed to fatty acid free BSA in a 5:1 molar ratio. Incubation with acompound and oleate is conducted for 24 hours. Media supernatants areremoved and ApoB concentrations are measured using ELISA kits (MabtechAB). Percent inhibition of ApoB secretion is determined by normalizingdata to vehicle treated wells. For a given compound, an IC₅₀(concentration at which 50% of ApoB secretion is inhibited) can also bedetermined by using a 4 parameter-fit inhibition curve model (Graph PadPrism®). In each experiment, cell viability is determined using theATPlite 1-Step kit (Perkin Elmer), such that compound effects due tocytotoxicity can be monitored.

The fatty acid niacin conjugate I-7 was evaluated in HepG2 cells at 3concentrations (50, 100 and 200 μM). The level of ApoB secretion wascompared to that of niacin, evaluated at 5 mM concentration. Compared toniacin, the fatty acid niacin conjugate I-7 showed significantinhibition of ApoB at a much lower drug concentration.

Example 2 Effect of Fatty Acid Niacin Derivatives on SREBP-1c TargetGenes

HepG2 cells (ATCC) were seeded at 20,000 cells per well in 96 wellplates. After adhering overnight, growth media (1.0% FBS in DMEM) wasremoved and cells were serum starved for 24 hours in DMEM containing 1%fatty acid free bovine serum albumin (BSA, Sigma). Cells were thentreated with one of four substances at a final concentration of 50 μM in1% BSA or 0.1% oleate complexed to fatty acid free BSA in a 5:1 molarratio (the four substances were compound I-7, compound I-8, acombination, of free niacin and free DHA, or a combination of freeniacin and free EPA). Cells were incubated for 6 hours and then washedwith PBS. RNA was reverse-transcribed using the cells to cDNA reagentsaccording to standard protocols (outlined in Applied Biosystem StepOneReal-time PCR protocols). Real time PCR of transcripts was performedwith Taqman assays for the three specific genes FASN (fatty acidsynthase), SCD (steroyl CoA desaturase) and ApoA1 (apolipoprotein A1).In all three cases, 18S-VIC® was used as a normalization control. Asshown in FIG. 2, statistically significant inhibition of FASN and SCDgene expression and an increase in ApoA1 gene expression were observedwhen HepG2 cells were stimulated with oleate in the presence of 50 μM ofcompound I-7 and compound I-8. The two groups containing a combinationof either free niacin and free DHA or niacin and free EPA produced nosignificant changes in the expression of these three specific genes at afinal concentration of 50 μM.

The activity of the niacin conjugated fatty acid mixtures can bedetermined using the above described assays.

Compounds

The following non-limiting compound examples serve to illustrate furtherembodiments of the fatty acid niacin derivatives. It is to be understoodthat any embodiments listed in the Examples section are embodiments ofthe fatty acid niacin derivatives and, as such, are suitable for use inthe methods and compositions described above.

Example 3 Preparation ofN-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)nicotinamide(I-7)

In a typical run, nicotinic acid (2.0 g, 16.2 mmol) was taken up inCH₂Cl₂ (20 mL) along with oxalyl chloride (1.4 mL, 16.2 mmol). After afew drops of DMF were added, the reaction mixture was stirred at roomtemperature until all the solids had dissolved and all gas evolution hadceased (1 h). This freshly prepared solution of the acid chloride wasadded dropwise at 0° C. to a solution containing tert-butyl2-aminoethylcarbamate (2.6 g, 16.2 mmol) and Et₃N (3.4 mL, 24.2 mmol) inCH₂Cl₂ (200 mL). The resulting reaction mixture was warmed to roomtemperature and stirred for 2 h. It was then washed with brine, driedover Na₂SO₄, filtered and concentrated under reduced pressure.Purification by silica gel chromatography (CH₂Cl₂) afforded tert-butyl2-(nicotinamido)ethyl carbamate (3.1 g, 74%).

tert-Butyl 2-(nicotinamido)ethylcarbamate (3.1 g, 11.7 mmol) was takenup in 25% TFA in CH₂Cl₂ (10 mL). The resulting reaction mixture wasallowed to stand at room temperature for 1 h. At this point, aconsiderable amount of precipitate formed and the clear filtrate wasremoved. The remaining solids were dried to afford of the TFA salt ofN-(2-aminoethyl)nicotinamide (1.6 g).

The TFA salt of N-(2-aminoethyl)nicotinamide (5.0 mmol) was taken up inCH₃CN (20 mL) along with(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (5.0mmol), HATU (5.5 mmol) and DIEA (15 mmol). The resulting reactionmixture was stirred at room temperature for 2 h and diluted with EtOAc.The organic layer was washed with saturated aqueous NaHCO₃, brine, driedover Na₂SO₄, filtered and concentrated under reduced pressure.Purification by silica gel chromatography (5% MeOH—CH₂Cl₂) affordedN-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)nicotinamide,calculated for C₃₀H₄₁N₃O₂: 475.32; found: [M+H]⁺476.

Example 4 Preparation ofN-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide(I-8)

The TFA salt of N-(2-aminoethyl)nicotinamide (1.6 g, 5.7 mmol) was takenup in CH₃CN (15 mL) along with(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoic acid (1.7 g, 5.7mmol), HATU (2.4 g, 6.3 mmol) and DIEA (3 mL, 17 mmol). The resultingreaction mixture was stirred at room temperature for 2 h and dilutedwith EtOAc. The organic layer was washed with saturated aqueous NaHCO₃,brine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. Purification by silica gel chromatography (5% MeOH—CH₂Cl₂)affordedN-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide(1.6 g, 62%), MS calculated for C₂₈H₃₉N₃O₂: 449.3; found: [M+H]⁺450.

Example 5 Preparation ofN-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)disulfanyl)ethyl)nicotinamide(I-3)

Cystamine dihydrochloride (1.0 g, 4.44 mmol) was dissolved in MeOH (50mL). Triethylamine (1.85 mL, 3 eq) was added at room temperature,followed by dropwise addition of Boc₂O (0.97 g, 4.44 mmol) as a solutionin MeOH (5 mL). The resulting reaction mixture was stirred at roomtemperature for 3 h. It was then concentrated under reduced pressure andthe resulting residue was taken up in 1M aqueous NaH₂PO₄ (20 mL). Theaqueous layer was washed with a 1:1 solution of pentane/EtOAc (10 mL),basified to pH 9 with 1M aqueous NaOH, and extracted with EtOAc. Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and concentrated under reduced pressure to afford tert-butyl2-(2-(2-aminoethyl)disulfanyl)ethylcarbamate (500 mg, 44%).

Separately, nicotinic acid (246 mg, 2.0 mmol) was taken up in CH₃CN (10mL) along with tert-butyl 2-(2-(2-aminoethyl)disulfanyl)ethylcarbamate(503 mg, 2.0 mmol), EDCI (422 mg, 2.2 mmol). The resulting reactionmixture was stirred at room temperature for 4 h and then diluted withEtOAc. The organic layer was washed with dilute aqueous NaHCO₃, brine,dried over Na₂SO₄, filtered and concentrated under reduced pressure.Purification by silica gel chromatography (CH₂Cl₂) afforded tert-butyl2-(2-(2-(nicotinamido)ethyl)disulfanyl)ethylcarbamate (400 mg, 56%).

tert-Butyl 2-(2-(2-(nicotinamido)ethyl)disulfanyl)ethylcarbamate (200mg, 0.56 mmol) was taken up in 25% TFA in CH₂Cl₂ solution (5 mL) andallowed to stand at room temperature for 4 h. The reaction mixture wasthen concentrated under reduced pressure to afford the TFA salt ofN-(2-(2-(2-aminoethyl)disulfanyl)ethyl)nicotinamide. This material wastaken up in CH₃CN (10 mL) along with(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (184 mg,0.56 mmol), HATU (234 mg, 0.62 mmol) and DIEA (0.30 mL). The resultingreaction mixture was stirred at room temperature for 2 h. It was thendiluted with EtOAc and washed successively with saturated aqueous NaHCO₃and brine. The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure. Purification by silica gelchromatography (5% MeOH—CH₂Cl₂) afforded(N-(2-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)disulfanyl)ethyl)nicotinamide(300 mg, 86%). MS calculated for C₃₂H₄₅N₃O₂S₂: 567.3; found: [M+H]⁺568.

Example 6 Preparation ofN-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethyl)nicotinamide(I-1)

In a typical run, sodium hydroxide (400 mg, 10 mmol) was dissolved inMeOH (79 mL) and 2-(2-aminoethoxy)ethanamine dihydrochloride (1.0 g,5.65 mmol) was added. The resulting reaction mixture was stirred at roomtemperature for 30 min. A solution containing Boc₂O (740 mg, 3.40 mmol)in THF (15 mL) was then added dropwise, at room temperature, over aperiod of 15 min. The resulting reaction mixture was stirred at roomtemperature for 18 h. It was then concentrated under reduced pressure.The resulting residue was taken up in CH₂Cl₂ (200 mL) and. stirredvigorously at room temperature for 4 h. The mixture was filtered and thefiltrate was concentrated under reduced pressure to afford tert-butyl2-(2-aminoethoxy)ethylcarbamate (850 mg, 74%).

tert-Butyl 2-(2-aminoethoxy)ethylcarbamate (420, 2.06 mmol was thentaken up in CH₃CN (20 ml) along with nicotinic acid (253 mg, 2.06 mmol)and EDCI (434 mg, 2.3 mmol). The resulting reaction mixture was stirredat room temperature for 15 h. It was then diluted with EtOAc (20 mL),washed with saturated aqueous NaHCO₃, brine, dried over Na₂SO₄, filteredand concentrated under reduced pressure. The resulting residue waspurified by silica gel chromatography (9:1 CH₂Cl₂/MeOH) to affordtert-butyl 2-(2-(nicotinamido)ethoxy)ethylcarbamate (280 mg, 44%). MScalculated for C₁₅H₂₃N₃O₄: 309.17; found: [M+H]⁺310.

tert-Butyl 2-(2-(nicotinamido)ethoxy)ethylcarbamate (140 mg, 0.453 mmol)was taken up in 25% TFA in CH₂Cl₂ (10 mL). The reaction mixture wasallowed to stand at room temperature for 2 h and then concentrated underreduced pressure to afford the TFA salt ofN-(2-(2-aminoethoxy)ethyl)nicotinamide. This material was then taken upin CH₃CN (10 mL) along with(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (148 mg,0.453 mmol), HATU (190 mg, 0.498 mmol) and DIEA (0.24 mL). The resultingreaction mixture was stirred at room temperature for 2 h. It was thendiluted with EtOAc and washed successively with saturated aqueous NaHCO₃and brine. The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure. Purification by silica gelchromatography (9:1 CH₂Cl₂/MeOH) affordedN-(2-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethoxy)ethyl)nicotinamide(75 mg, 31%). MS calculated for C₃₁H₄₆N₂O₅: 526.34; found: [M+H]⁺527.

Example 7 Preparation ofN-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(methyl)amino)ethyl)nicotinamide(I-2)

N1-(2-Aminoethyl)-N1-methylethane-1,2-diamine (5.0 g, 42.7 mmol) wasdissolved in CH₂Cl₂ (100 mL) and cooled to 0° C. A solution of Boc₂O(0.93 g, 4.27 mmol) in CH₂Cl₂ (10 mL) was then added dropwise at 0° C.over a period of 15 min. The resulting reaction mixture was stirred at0° C. for 30 min and then warmed to room temperature. After stirring atroom temperature for 2 h, the reaction mixture was diluted with CH₂Cl₂(100 mL). The organic layer was washed with brine (3×25 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to affordtert-butyl 2-((2-aminoethyl)(methyl)amino)ethylcarbamate (1.1 g).

tert-Butyl 2-((2-aminoethyl)(methyl)amino)ethylcarbamate (400 mg, 1.84mmol) was taken up in CH₃CN (10 mL) along with nicotinic acid (227 mg,1.84 mmol) and EDCI (353 mg, 2.02 mmol). The resulting reaction mixturewas stirred at room temperature for 18 h and then diluted with EtOAc.The organic layer was washed with saturated aqueous NaHCO₃, brine, driedover Na₂SO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by silica gel chromatography (5%MeOH—CH₂Cl₂) to afford tert-butyl2-(methyl(2-(nicotinamido)ethyl)amino)ethylcarbamate (180 mg, 30%). MScalculated for C₁₆H₂₆N₄O₃: 322.2; found: [M+H]⁺323.

tert-Butyl 2-(methyl(2-(nicotinamido)ethyl)amino)ethylcarbamate (90 mg,0.279 mmol) was taken up in a 25% TFA in CH₂Cl₂ solution (5 mL) andallowed to stand at room temperature for 3 h. The reaction mixture wasconcentrated under reduced pressure to afford the TFA salt ofN-(2-((2-aminoethyl)(methyl)amino)ethyl)nicotinamide. This material wastaken up in CH₃CN (10 mL) along with(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (90 mg,0.279 mmol), HATU (117 mg, 0.31 mmol) and DIEA (0.15 mL). The resultingreaction mixture was stirred at room temperature for 2 h. It was thendiluted with EtOAc and washed successively with saturated aqueous NaHCO₃and brine. The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure. Purification by silica gelchromatography (5% MeOH—CH₂Cl₂) affordedN-(2-((2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)(methyl)amino)ethyl)nicotinamide(30 mg, 20%). MS calculated for C₃₃H₄₈N₄O₂: 532.38; found: [M+H]⁺533.

Example 8 Preparation of (2S,3R)-methyl3-((S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoyloxy)-2-(nicotinamido)butanoate(I-9)

L-Alanine methyl ester hydrochloride (0.85 g, 6.1 mmol) was taken up inCH₃CN (20 mL) along with(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (2.0 g,6.1 mmol, EDCI (1.3 g, 6.72 mmol) and DIEA (1.3 mL). The resultingreaction mixture was stirred at room temperature for 2 h. It was thendiluted with EtOAc and washed with dilute aqueous NaHCO₃ and brine. Theorganic layer was dried over Na₂SO₄, filtered and concentrated underreduced pressure to afford (S)-methyl2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoate(2.0 g, 79%).

(S)-methyl2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoate(2.0 g, 4.8 mmol) was taken up in THF (8 mL) along with 5M aqueous NaOH(5 mL) and stirred vigorously at room temperature for 3 h. The reactionmixture was diluted with water and concentrated under reduced pressure.Enough 6N HCl was then added to adjust the pH to 2. The resultingmixture was extracted with EtOAc. The combined organic layers were driedover Na₂SO₄, filtered and concentrated under reduced pressure to afford(S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoicacid. This was taken up in CH₃CN (15 mL) along with N-Boc-L-threoninemethyl ester (1.11 g, 4.78 mmol), HATU (2.0 g, 5.3 mmol) and DIEA (1.2mL). The resulting reaction mixture was stirred at room temperature for6 h and diluted with EtOAc. The organic layer was washed with NaHCO₃,brine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. Purification by silica gel chromatography (CH₂Cl₂) afforded(2S,3S)-methyl2-(tert-butoxycarbonyl)-3-((S)-2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoyloxy)butanoate(1.0 g).

(2S,3R)-methyl2-(tert-butoxycarbonyl)-3-((S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoyloxy)butanoate(300 mg, 0.488 mmol) was taken up in 4M HCl in dioxane (2 mL) andallowed to stand at room temperature for 10 min. The reaction mixturewas then diluted with EtOAc and concentrated under reduced pressure toafford the HCl salt of (2S,3R)-methyl2-amino-3-((S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoyloxy)butanoate.This material was taken up in CH₃CN (5 mL) along with nicotinic acid (60mg, 0.488 mmol), HATU (204 mg, 0.54 mmol) and DIEA (0.25 mL, 1.46 mmol).The resulting reaction mixture was stirred at room temperature for 1 hand concentrated under reduced pressure. The resulting oily residue waspurified by silica gel chromatography (9:1 CH₂Cl₂/MeOH) to afford(2S,3R)-methyl3-((S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoyloxy)-2-(nicotinamido)butanoate(120 mg, 40%). MS calculated for C₃₆H₄₉N₃O₆: 619.36; found: [M+H]⁺620.

Example 9 Preparation of (2S,3R)-methyl3-((S)-2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenamido)propanoyloxy)-2-(nicotinamido)butanoate(I-10)

The same synthetic sequence outlined above for the preparation of(2S,3R)-methyl3-((S)-2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)propanoyloxy)-2-(nicotinamido)butanoatewas used, except that (5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoicacid (EPA) was used instead of DHA. MS calculated for C₃₄H₄₇N₃O₆:593.35; found: [M+H]⁺594.

Example 10 Preparation of (S)-methyl6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)hexanoate(I-11)

H-Lysine-(BOC)-OMe hydrochloride (500 mg, 1.68 mmol) was taken up inCH₃CN (10 mL) along with nicotinic acid (207 mg, 1.68 mmol), EDCI (354mg, 1.85 mmol) and DIEA (0.90 mL). The resulting reaction mixture wasstirred at room temperature for 18 h and diluted with EtOAc. The organiclayer was washed with dilute aqueous NaHCO₃, brine, dried over Na₂SO₄,filtered and concentrated under reduced pressure. Purification by silicagel chromatography (CH₂Cl₂) afforded (S)-methyl6-(tert-butoxycarbonyl)-2-(nicotinamido)hexanoate (520 mg, 85%).

(S)-Methyl 6-(tert-butoxycarbonyl)-2-(nicotinamido)hexanoate (260 mg,0.71 mmol) was taken up in 4M HCl in dioxane (2 mL) and allowed to standat room temperature for 1 h. The reaction mixture was diluted with EtOAcand concentrated under reduced pressure to afford the HCl salt of(S)-methyl 6-amino-2-(nicotinamido)hexanoate. This material was taken upin CH₃CN (5 mL) along with(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (233 mg,0.71 mmol), HATU (297 mg, 0.78 mmol) and DIEA (0.4 mL). The resultingreaction mixture was stirred at room temperature for 2 h and dilutedwith EtOAc. The organic layer was washed with dilute aqueous NaHCO₃,brine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. Purification by silica gel chromatography (9:1 CH₂Cl₂/MeOH)afforded (S)-methyl6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)hexanoate(280 mg, 72%). MS calculated for C₃₅H₄₉N₃O₄: 575.37; found: [M+H]⁺576.

Example 11 Preparation of(S)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)hexanoicacid (I-12)

(S)-Methyl6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)hexanoate(40 mg, 0.0695 mmol) was taken up in 2 mL of THF along with 80 μL of a 5M NaOH solution. The resulting reaction mixture was stirred at roomtemperature for 2 h. It was then acidified to pH 4 with 2 N HCl and thenextracted with EtOAc. The combined organic layers were dried (Na₂SO₄)and concentrated under reduced pressure to afford 31 mg of(S)-6-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)-2-(nicotinamido)hexanoicacid. MS calculated for C₃₄H₄₇N₃O₄: 561.36; found: [M+H]⁺562.

Example 12 Preparation of (S)-methyl2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-6-(nicotinamido)hexanoate(I-13)

H-Lysine-(BOC)-OMe hydrochloride (500 mg, 1.68 mmol) was taken up in 25mL of CH₃CN along with(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoic acid (EPA, 509 mg,1.68 mmol), HATU (702 mg, 1.85 mmol) and DIEA (880 μL, 5.04 mmol). Theresulting reaction mixture wax stirred at room temperature for 2 h. Itwas then diluted with EtOAc (70 mL) and washed with brine (20 mL). Theorganic layer was dried (Na₂SO₄) and concentrated under reducedpressure. The resulting residue was purified by silica gelchromatography (CH₂Cl₂, gradient elution to 90% CH₂Cl₂, 10% MeOH) toafford 870 mg of (S)-methyl6-(tert-butoxycarbonyl)-2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)hexanoate(95% yield). MS calculated for C₃₂H₅₂N₂O₅: 544.39; found: [M+H]⁺545.

(S)-Methyl6-(tert-butoxycarbonyl)-2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)hexanoate(870 mg, 1.60 mmol) was taken up in 4 mL of 4 N HCl in dioxane andallowed to stand at room temperature for 10 min. The reaction mixturewas diluted with 10 mL of EtOAc and concentrated under reduced pressureto afford the HCl salt of (S)-methyl6-amino-2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)hexanoate.This residue was taken up in 5 mL of CH₃CN along with nicotinic acid(197 mg, 1.60 mmol), HATU (669 mg, 1.76 mmol) and DIEA (836 mL, 4.8mmol). The resulting reaction mixture was stirred at room temperaturefor 2 h and diluted with EtOAc (20 mL). The organic layer was washedwith brine (20 mL), dried (Na₂SO₄) and concentrated under reducedpressure. The resulting residue was purified by chromatography (95%CH₂Cl₂, 5% MeOH) to afford 300 mg of (S)-methyl2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-6-(nicotinamido)hexanoate.MS calculated for C₃₃H₄₇N₃O₄: 549.36; found: [M+H]⁺550.

Example 13 Preparation of(S)-2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-6-(nicotinamido)hexanoicacid (I-14)

(S)-methyl2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-6-(nicotinamido)hexanoate(140 mg, 0.225 mmol) was taken up in 2 mL of THF along with an aqueoussolution of NaOH (35 mg in 2 mL of H₂O). The resulting reaction mixturewas stirred at room temperature for 2 h. It was then acidified to pH 4with 2 N HCl and then extracted with EtOAc. The combined organic layerswere dried (Na₂SO₄) and concentrated under reduced pressure to afford 31mg of(S)-2-((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamido)-6-(nicotinamido)hexanoicacid. MS calculated for C₃₄H₄₇N₃O₄: 561.36; found: [M+H]⁺562. MScalculated for C₃₂H₄₅N₃O₄: 535.34; found: [M+H]⁺536.

Mixtures Synthesis of a mixture ofN-(2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)ethyl)nicotinamideandN-(2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenamido)ethyl)nicotinamide

A 1.27:1 mixture of (4Z,7Z,10Z,13Z,16Z,19Z)-ethyldocosa-4,7,10,13,16,19-hexaenoate and (5Z,8Z,11Z,14Z,17Z)-ethylicosa-5,8,11,14,17-pentaenoate was taken up in 130 mL of THF and 110 mLof a 5 N NaOH solution was added. The resulting reaction mixture wasstirred at 50° C. for 4 h. It was then cooled to room temperature andconcentrated under reduced pressure. The resulting aqueous mixture wascooled in an ice bath, acidified to pH 2 with 6 N HCl and extracted withEtOAc (3×200 mL). The combined organic layers were washed with water(5×200 mL), brine (300 mL), dried (Na₂SO₄) and concentrated underreduced pressure to afford 85 g of a 1.27:1 mixture of(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid and(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoic acid. The HCl salt ofN-(2-aminoethyl)nicotinamide was prepared according to the procedureoutlined in US 20110053990. This material (9.4 g, 39.66 mmol) was takenup in 150 mL of CH₂Cl₂ along with 10 g of the 1.27:1 mixture of(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid and(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoic acid, EDC (7.0 g,36.4 mmol), HOBT (4.9 g, 36.4 mmol) and Et₃N (14 mL, 99.3 mmol). Theresulting reaction mixture was stirred at room temperature for 18 h. Itwas then washed with saturated NH₄Cl (150 mL), brine (150 mL), dried(Na₂SO₄) and concentrated under reduced pressure. The resulting productwas purified by silica gel chromatography (95% CH₂Cl₂, 5% MeOH) toafford 13.2 g of the 1.27:1 mixture ofN-(2-((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamido)ethyl)nicotinamideandN-(2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenamido)ethyl)nicotinamide.MS (ES⁺) calculated for C₃₀H₄N₃O₂ and C₂₈H₃₉N₃O₂: 475.32 and 449.30respectively; found 476 and 450 respectively.

The present invention is not to be limited in scope by the specificembodiment disclosed in the examples which are intended as illustrationsof a few aspects of the invention and any embodiments that arefunctionally equivalent are within the scope of this invention. Indeed,various modifications of the invention in addition to those shown anddescribed herein will become apparent to those skilled in the art andare intended to fall within the scope of the appended claims.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain, usingno more than routine experimentation, numerous equivalents to thespecific embodiments described specifically herein. Such equivalents areintended to be encompassed in the scope of the following claims.

1-10. (canceled)
 11. A method for treating or preventing a metabolicdisorder comprising the administration of an effective amount of acomposition comprising a mixture of a niacin-EPA conjugate and aniacin-DHA conjugate wherein a niacin is linked to either DHA or EPA viaa linker having the formula:

wherein, each a, b, c, and d is independently —H, -D, —CH₃m, —OCH₃,—OCH₂CH₃, —C(O)OR, or benzyl; up to two of a, b, c, and d can be —OZ; ortwo of a, b, c, and d can be taken together, along with the singlecarbon to which they are bound, to form a cycloalkyl or heterocycle; mis 0, 1, 2, or 3; each n, o, p, and q is independently 0 or 1; each L isindependently —O—, —S—, —S(O)—, —S(O)₂—, —S—S—,

wherein when m is 2 or 3, up to one of the heteroatoms —O—, —S—, —S(O)—,—S(O)₂—, —S—S— can be selected for L; each g is independently 2, 3 or 4;each h is independently 1, 2, 3 or 4; each R₆ is independently H orC₁-C₆ alkyl, or both groups, when taken together with the nitrogen towhich they are attached, form a heterocycle; each R₇ is independently e,H, or straight or branched C₁-C₁₀ alkyl which can be optionallysubstituted with OH, NH₂, CO₂R, CONH₂, phenyl, C₆H₄OH, imidazole orarginine; each e is independently H or any one of the side chains of anaturally occurring amino acid; each R is independently —H, —C(O)—C₁-C₃alkyl, or straight or branched C₁-C₄ alkyl optionally substituted withhalogen; wherein Z is independently —H, or

each r is independently 2, 3, or 7; each s is independently 3, 5, or 6;each t is independently 0 or 1; each v is independently 1, 2, or 6; andR₄ and R₅ are each independently —H, -D, —C₁-C₄ alkyl, -halogen, —OH,—C(O)C₁-C₄ alkyl, —O-aryl, —O-benzyl, —OC(O)C₁-C₄ alkyl, —C₂-C₃ alkene,—C₂-C₃ alkyne, —NH₂, —NH(C₁-C₃ alkyl), —N(C₁-C₃ alkyl)₂, —NH(C(O)C₁-C₃alkyl), —N(C(O)C₁-C₃ alkyl)₂, —SH, —S(C₁-C₃ alkyl), —S(O)C₁-C₃ alkyl, or—S(O)₂C₁-C₃ alkyl.
 12. The method of claim 11, wherein the niacin-EPAconjugate and the niacin-DHA conjugate comprise:N-(2-(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenamidoethyl)nicotinamide(I-7); andN-(2-(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenamidoethyl)nicotinamide(I-8).
 13. The method of claim 11, wherein the EPA is EPA ethyl esterand the DHA is DHA ethyl ester.
 14. The method of claim 11, wherein theEPA and DHA are in the form of free acids.
 15. The method of claim 11,wherein the ratio of EPA to DHA is between about 25:75 to about 75:25.16. The method of claim 11, wherein the linker is ethylenediamine. 17.The method of claim 11, wherein the metabolic disorder isatherosclerosis, dyslipidemia, hypertriglyceridemia, hypertension, heartfailure, cardiac arrhythmias, low HDL levels, high LDL levels, suddendeath, stable angina, coronary heart disease, acute myocardialinfarction, secondary prevention of myocardial infarction,cardiomyopathy, endocarditis, type 2 diabetes, insulin resistance,impaired glucose tolerance, hypercholesterolemia, stroke,hyperlipidemia, hyperlipoproteinemia, chronic kidney disease,intermittent claudication, hyperphosphatemia, carotid atherosclerosis,peripheral arterial disease, diabetic nephropathy, hypercholesterolemiain HIV infection, acute coronary syndrome (ACS), non-alcoholic fattyliver disease, arterial occlusive diseases, cerebral arteriosclerosis,cerebrovascular disorders, myocardial ischemia, and diabetic autonomicneuropathy.